44 NATURALS NT NS = not treated not stimulated, NT UV = not treated UV irradiated 2.5
* +
2 +61% 1.33 1.5 1.07 1 0.5 0 NT NS NT UV 0.01% 0.001% Vitamin B12
Study 1: Could Vitamin B12 reduce skin inflammatory reactions caused by inflammasome activation? Inflammasomes are intracellular, multi-protein complexes that form part of our innate immune response, playing a defensive role against pathogens. In skin, UV radiation, pollution and oxidative stress induce the formation of the NLRP3- inflammasome in keratinocytes in the epidermis. NLRP3-inflammasome is composed of the
NLRP3 pyrin, ASC (apoptosis associated speck containing a caspase activation and recruitment domain), and the inactive protease enzyme Pro-caspase-1. It is responsible for regulating the immunological response and triggers an inflammatory response by producing cytokines such as IL-1 β and IL-18. Chronic activation of NLRP3-inflammasome can lead to skin irritation, symptoms such as redness, irritation resulting in skin sensitivity, and signs of premature skin ageing. Earlier studies have shown that NLRP3- inflammasome is regulated by an epigenetic factor such as the micro-RNA, miR-22-3-p.5 Figure 1 illustrates the overall mechanism of action. In the first of our new studies, we investigated the action of Vitamin B 12 on epigenetic factor
miR-22-3-p as negative regulator of inflammasome and its involved downstream activation targets, ASC complex and pro-IL-1β. With this, B12 indeed showed upregulation of hsa-miR-22-3p.
Methodology and findings For our study, we investigated the effects of vitamin B12 at a concentration of 0.01% on a human skin explant model exposed to UVB irradiation. UVB was used as a stress factor to stimulate inflammasome activation. To do this, having first evaluated the cytotoxicity of the vitamin B12, we quantified IL-1β in a culture medium using an ELISA kit assay. Then, we used immunohistochemistry to study
the effect of the active ingredient on ASC complex activation. Finally, from quantitative reverse transcription PCR (RT-qPCR), we quantified the hsa-miR-22-3p regulating the inflammasome. Our findings are illustrated in the graphs in Figure 2. The graph on the left represents expression of
the epigenetic factor miR-22-3p after UV stress for a sample treated with 0.01% and 0.001% vitamin B12 and for control samples (untreated and/or not irradiated). It shows that at a concentration of 0.01%, vitamin B12 significantly upregulated miR-
+24% 1.72
160 140 120 100 80 60 40 20 0
+++ 135
100 *** vs NT NS p<0.001 +++vs NT UV p<0.001 -100% +++ 100 -46% +++
119 ***
NT NS
NT UV
0.01% Figure 2: Effects of Vitamin B12 on miR-22-3p expression after UV stress (left) and on ASC complex activation (right) after UVB stress
22-3p by 61% compared to the untreated samples. The graph on the right represents the mean
ASC intensity for the same sample and after UVB stress. This shows that at 0.01%, Vitamin B12 significantly downregulated ASC complex activation by 100%, i.e., to the level of a sample that had not been exposed to UVB irradiation. To finish this study, we also evaluated the
effect of vitamin B12 treatment on IL-1β production after UVB stress and found that at 0.01%, the vitamin decreased production of this pro- inflammatory cytokine by 35%. Overall, these ex vivo findings give an
encouraging indication that by reducing inflammasome, vitamin B12 could help relieve symptoms of skin sensitivity induced by UV exposure and other external stressors.
Study 2: Could vitamin B12 reduce skin sensitivity and inflammation caused by mast cell degranulation? Mast cells are immune cells present in most tissues but particularly prominent in organs exposed to the environment such as the skin. They can be activated by environmental stress factors like allergens, UV light, pollution and
0.001% Vitamin B12
Figure 3: Mechanism of action for mast cell degranulation and the inhibition of histamine release PERSONAL CARE October 2022
www.personalcaremagazine.com
miR-22-3p expression after UV stress
Mean ASC intensity
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